LOCUS       sequence-215501-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 223 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3040)
  AUTHORS   Taylor GM, Mordaka PM, Heap JT
  TITLE     Start-Stop Assembly: a functionally scarless DNA assembly system 
            optimized for metabolic engineering.
  JOURNAL   Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 
            10.1093/nar/gky1182.
  PUBMED    30462270
REFERENCE   2  (bases 1 to 3040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2018 Nov 20. pii: 5193345. doi: 10.1093/nar/gky1182."
COMMENT     Sequence Label: pStA223
FEATURES             Location/Qualifiers
     source          1..3040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..95
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(181..186,187..204,205..211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    219..235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     224..246
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     243..259
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     243..259
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             255..578
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    272..328
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(329..346)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(329..345)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(338..360)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(554..573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(607..693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1043..1588
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1489..1508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1705..1722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2172..2191)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2183..2998
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2183..2998
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2256..2275)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
       61 tcggttgccg ccgggcgttt tttattggtg agaatgaaga caaactagga gagagaccca
      121 gtagggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      181 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
      301 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
      361 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      421 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      481 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      541 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt tccggtctca tactttacaa
      601 gtcttcattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      661 ccagtctttc gactgagcct ttcgttttat ttgatgcccg aaagggcctc gtgatacgca
      721 aatctggagc cggtgagcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      781 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      841 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
      901 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
      961 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1021 accccttaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     1081 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     1141 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     1201 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     1261 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     1321 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     1381 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     1441 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     1501 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     1561 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     1621 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     1681 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     1741 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     1801 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgagg
     1861 tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag
     1921 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat
     1981 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc
     2041 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg
     2101 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2161 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc
     2221 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2281 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     2341 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2401 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2461 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     2521 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2581 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2641 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2701 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     2761 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2821 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2881 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2941 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3001 ttgggggaaa ttagccgcag ccgaacgacc gagcgccttg
//
